This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Metabolomics is the unbiased study of small molecules in biological samples. We have previously generated methods for targeted metabolite profiling of amino acids and other classes of small molecules in yeast. Now we have focused our efforts on using two-dimensional gas chromatography with mass spectrometry for an untargeted metabolomics approach in yeast. We have optimized these protocols for the quantification of approximately 150-200 different small molecule metabolites found in yeast. We are applying these methods for the characterization of natural variation in wild yeast strains. In collaboration with other members of the yeast resource center, we are measuring metabolic profiles for 38 individual strains grown under chemostat controlled conditions. Our data in combination with gene expression and proteomics data could provide a more complete network of gene regulation, gene expression and cellular metabolism differ among these strains. This project is just one example of the applications for this technology which can easily be applied to human biofluids and tissue culture samples.